Abstract
The characteristics of tongue coating are very important symbols for disease diagnosis in traditional Chinese medicine (TCM) theory. As a habitat of oral microbiota, bacteria on the tongue dorsum have been proved to be the cause of many oral diseases. The high-throughput next-generation sequencing (NGS) platforms have been widely applied in the analysis of bacterial 16S rRNA gene. We developed a methodology based on genus-specific multiprimer amplification and ligation-based sequencing for microbiota analysis. In order to validate the efficiency of the approach, we thoroughly analyzed six tongue coating samples from lung cancer patients with different TCM types, and more than 600 genera of bacteria were detected by this platform. The results showed that ligation-based parallel sequencing combined with enzyme digestion and multiamplification could expand the effective length of sequencing reads and could be applied in the microbiota analysis.
Highlights
The complex microbial flora living on or within the human body has long been proposed to contribute to the human health as well as disease [1–8] (Eckburg et al, 2005)
Since SOLiD system employs 2-base encoding and color-space strategy, a single color change is a measurement error, two adjacent color changes may result from a single nucleotide variation in base space, and three color-space mismatches might imply two adjacent variants compared to reference
We explored a method to detect the microbial diversity using 16S rRNA gene by high-throughput SOLiD sequencing system
Summary
The complex microbial flora living on or within the human body has long been proposed to contribute to the human health as well as disease [1–8] (Eckburg et al, 2005). As a reservoir for oral microorganisms, food scraps, saliva, and shed epithelial cells, the human tongue has been investigated showing significant association with the microbial communities of the gut and diseases such as gastritis or halitosis [11, 21–24]. Han et al sequenced the V2–V4 region of 16S rRNA gene by pyrosequencing to investigate the tongue coating microbiome in patients with colorectal cancer and healthy controls. Their results indicated that the richness of the bacterial communities in the patients with thin tongue coating and healthy controls was higher than in the patients with thick tongue coating
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